Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 98, Issue 3, pp 331–340

Evaluation of parameters affecting Agrobacterium-mediated transformation of citrus

Original Paper

DOI: 10.1007/s11240-009-9567-1

Cite this article as:
Dutt, M. & Grosser, J.W. Plant Cell Tiss Organ Cult (2009) 98: 331. doi:10.1007/s11240-009-9567-1

Abstract

An improved protocol for genetic transformation of juvenile explants of Carrizo (Citrus sinensis Osb. × Poncirus trifoliata L. Raf.), Duncan (Citrus paradisi Macf.), Hamlin (Citrus sinensis (L.) Osbeck) and Mexican Lime (Citrus aurantifolia Swingle) cultivars using a vector containing a bifunctional egfp-nptII fusion gene is described. Several parameters were investigated to optimize genetic transformation of these four cultivars. It was determined that a short preincubation in hormone rich liquid medium and subculture of Agrobacterium for 3 h in YEP medium containing 100 μM acetosyringone were required for improvement of transformation efficiency. Co-cultivation duration as well as addition of acetosyringone to co-cultivation medium also played an important role in transformation efficiency as did OD600 value of the Agrobacterium suspension used for transformation. We regenerated numerous EGFP expressing transgenic lines from all four cultivars. Based on these results, we conclude that genetic transformation of citrus is cultivar specific and optimization of conditions for maximum transgenic production are required for each individual cultivar.

Keywords

Agrobacterium tumefaciens Bifunctional gene Citrus EGFP Transformation 

Abbreviations

BAP

Benzyladenine

EGFP

Enhanced green fluorescent protein

NAA

Napthyleneacetic acid

YEP

Yeast extract peptone

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  1. 1.Horticultural Sciences Department, Citrus Research and Education Center (CREC)University of Florida-IFASLake AlfredUSA

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